Spiro-substituted pyrrolopyrimidines

ABSTRACT

The invention provides compounds of formula I or a pharmaceutically acceptable salt or ester thereof formula I 
     
       
         
         
             
             
         
       
     
     wherein the symbols have the meaning as defined in the description. Said compounds are inhibitors of cathepsin K and/or cathepsin S and are useful for the treatment of diseases and medical conditions in which cathepsin K and or cathepsin S is implicated, e.g. various disorders including neuropathic pain, inflammation, rheumatoid arthritis, osteoarthritis, osteoporosis, multiple sclerosis and tumours.

This invention relates to inhibitors of cysteine proteases, inparticular to pyrrolopyrimide cathepsin K inhibitors or cathepsin Sinhibitors or inhibitors with mixed activities and to theirpharmaceutical use for the treatment or prophylaxis of diseases ormedical conditions in which cathepsin K or cathepsin S is implicated orboth are implicated.

Cathepsin K and cathepsin S are members of the family of lysosomalcysteine cathepsin enzymes, comprising e.g. cathepsins B, K, L and S,which are implicated in various disorders including neuropathic pain,inflammation, rheumatoid arthritis, osteoarthritis, osteoporosis, tumors(especially tumor invasion and tumor metastasis), obesity, coronarydisease, atherosclerosis (including atherosclerotic plaque rupture anddestabilization), autoimmune diseases, multiple sclerosis, respiratorydiseases, infectious diseases and immunologically mediated diseases(including transplant rejection). Thus compounds of the invention whichare dual inhibitors to cathepsin K and cathepsin S or specificinhibitors to cathepsin S or cathepsin K may be useful in theherein-mentioned diseases.

Accordingly the present invention provides a compound of formula I, or apharmaceutically acceptable salt or ester thereof

wherein E is a radical of formula a or formula b

wherein

A is CH₂, CH₂—CH₂ or C═O; B is CH₂, C═O or

D is CH₂, or C═O; G is CH₂, CH₂C═O or CH₂—CH₂; J is CH₂, C═O or CH₂—CH₂;

L is H, OCH₃, halo, or lower alkoxy;

M is CH₂ or NH;

Q is H, lower alkyl, hydroxy substituted lower alkyl, optionallysubstituted aryl lower alkyl, lower alkyl sulfonyl, carbocyclic aryllower alkyl, lower alkoxy-substituted carbocyclic aryl lower alkyl,halo-substituted carbocyclic aryl lower alkyl,N-heterocyclyl-substituted lower alkyl, lower alkoxy substitutedcarbocyclic aryl, amino carbonyl, cycloalkyl amino carbonyl,N-heterocyclyl substituted lower alkyl carbonyl, halo-substitutedcarbocyclic aryl lower alkyl, lower alkoxy carbonyl, or lower alkylcarbonyl; andR is lower alkyl, para-chlorophenylethyl, cyclohexylethyl,dimethylbutyl, difluorocyclohexylethyl, cyclopentylethyl orcycloheptylethyl.

In a preferred embodiment of the invention provides a compound offormula I-(i), or a pharmaceutically acceptable salt or ester thereof

whereinQ-i is H, lower alkyl, hydroxyl-substituted lower al 1, N-heterocyclylsubstituted lower al 1, mono or di-substituted aryl lower alkyl, loweralkoxy substituted carbocyclic aryl lower alkyl; andR is as defined above.

In another preferred embodiment of the invention provides A compound offormula I-(ii), or a pharmaceutically acceptable salt or ester thereof

whereinQ-ii is H, lower alkyl, N-heterocyclyl substituted lower alkyl, halosubstituted carbocyclic aryl lower alkyl, lower alkyl carbonyl; andL and R are as defined above.

In a further preferred embodiment of the invention provides a compoundof formula I-(iii), or a pharmaceutically acceptable salt or esterthereof

wherein

B-iii is CH₂; G-iii is CH₂CH₂; J-iii is CH₂CH₂;

Q-iii is H, cycloalkyl amino carbonyl, amino carbonyl, lower alkoxysubstituted carbocyclic aryl, lower alkyl carbonyl, carbocyclic aryllower alkyl or N-heterocyclyl substituted lower alkyl carbonyl; andR is as defined above.

In a further preferred embodiment of the invention provides a compoundof formula I, formula I (i), formula I-(ii) or formula I-(iii), whereinR is R1=lower alkyl.

In a further preferred embodiment of the invention provides a compoundof formula I, formula I-(i), formula I-(ii) or formula I-(iii), whereinR is R5=2,2-dimethyl-propyl.

In a further preferred embodiment of the invention provides a compoundof formula I, formula I-(i), formula I-(ii) or formula I-(iii), whereinR is R6=3,3-dimethyl-butyl.

In a further preferred embodiment of the invention provides a compoundof formula I, formula I-(i), formula I-(ii) or formula I-(iii), whereinR is R2=para-chlorophenylethyl, cyclohexylethyl, dimethylbutyl,difluorocyclohexylethyl, cyclopentylethyl or cycloheptylethyl.

In a further preferred embodiment of the invention provides a compoundof formula I-(iv), or a pharmaceutically acceptable salt or esterthereof

whereinR3 is (8-lower alkyl-carbonyl)-2,8-diaza-spiro[4.5]dec-2-ylmethyl,(8-lower alkyl-sulfonyl)-2,8-diaza-spiro[4.5]dec-2-ylmethyl,(8-aryl-lower alkyl)-2,8-diaza-spiro[4.5]dec-2-ylmethyl,

R4 is para-chlorophenylmethyl, cyclohexylmethyl, dimethylpropyl,difluorocyclohexylmethyl, cyclopentylmethyl or cycloheptylmethyl; andQ is as defined above.

The present invention further provides processes for the preparation ofcompounds of formula I and their salts and esters, comprising the stepof coupling a compound of formula II

wherein Q, G, J, M, A, B, D are as defined above, with a compound offormula III

wherein X is a halo and R is defined above, and recovering the resultingcompound in free base, or in a pharmaceutically acceptable salt or esterthereof.

The above coupling procedure may be carried out in solution e.g. DMFsolution in the presence of K₂CO₃, for instance at room temperature withstirring e.g. for about 12 hours. As appropriate protecting groups maybe used to protect reactive functional groups during the couplingprocedure and may be removed after the coupling procedure, for instanceas hereinafter described in the Examples.

Working up the reaction mixtures and purification of the compounds thusobtained may be carried out in accordance to known procedures oraccording to the Examples.

Above and elsewhere in the present description the following terms havethe following meanings. Halo or halogen denote I, Br, Cl or F.

The term “lower” referred to above and hereinafter in connection withorganic radicals or compounds respectively defines such as branched orunbranched with up to and including 6 carbon atoms.

A “lower alkyl” group is branched or unbranched and contains 1 to 6carbon atoms. Lower alkyl represents, for example, methyl, ethyl,propyl, butyl, isopropyl isobutyl, tertiary butyl or neopentyl(2,2-dimethylpropyl).

“Halo-substituted lower alkyl” is C1-C7 lower alkyl substituted by up to6 halo atoms, preferably mono, di or tri-substituted lower alkyl.

A “lower alkoxy” group is branched or unbranched and contains 1 to 6carbon atoms, preferably 1-4 carbon atoms. Lower alkoxy represents forexample methoxy, ethoxy, propoxy, butoxy, isopropoxy, isobutoxy ortertiary butoxy.

“Aryl” represents a phenyl or naphthyl radical. Preferably a“carbocyclic aryl” consisting solely of carbon and hydrogen atomsoptionally substituted, mono-, di- or tri-substituted by one, two orthree radicals selected from lower alkyl, lower alkoxy, hydroxy,halogen, Preferred as carbocyclic aryl is phenyl or phenyl optionallysubstituted, for instance, as described in the examples, e.g. mono-, di-or tri-substituted by halogen, lower alkyl, lower or alkoxy.

“Cycloalkyl” represents a saturated cyclic hydrocarbon optionallysubstituted by lower alkyl which contains 3 to 10 ring carbons and isadvantageously cyclopropyl, cyclopentyl, cyclohexyl or optionallysubstituted by lower alkyl.

“N-heterocyclyl” represents a saturated, partially unsaturated oraromatic nitrogen containing heterocyclic moiety attached via a nitrogenatom having from 3 to 8 ring atoms, optionally containing a further Oheteroatom optionally substituted by a lower alkyl or lower alkylcarbonyl.

“Lower alkyl carbonyl” refers to a radical of the formula —C(O)R_(a)where R_(a) is a lower alkyl radical defined above, for example, acetyl,ethylcarbonyl, or n-propylcarbonyl.

Compounds of the invention are either obtained in the free form, or as asalt thereof if salt forming groups are present. Compounds of theinvention having basic groups can be converted into acid addition salts,especially pharmaceutically acceptable salts. These are formed, forexample, with inorganic acids, such as mineral acids, for examplesulfuric acid, a phosphoric or hydrohalic acid, or with organiccarboxylic acids, such as (C1-C4) alkanecarboxylic acids which, forexample, are unsubstituted or substituted by halogen, for example aceticacid, such as saturated or unsaturated dicarboxylic acids, for exampleoxalic, succinic, maleic or fumaric acid, such as hydroxycarboxylicacids, for example glycolic, lactic, malic, tartaric or citric acid,such as amino acids, for example aspartic or glutamic acid, or withorganic sulfonic acids, such as (C1-C4)-alkylsulfonic acids (for examplemethanesulfonic acid) or arylsulfonic acids which are unsubstituted orsubstituted (for example by halogen). Preferred are salts formed withhydrochloric acid, methanesulfonic acid and maleic acid.

In view of the close relationship between the free compounds and thecompounds in the form of their salts, whenever a compound is referred toin this context, a corresponding salt is also intended, provided such ispossible or appropriate under the circumstances.

The compounds, including their salts, can also be obtained in the formof their hydrates, or include other solvents used for theircrystallization.

If one or more other functional groups, for example carboxy, hydroxy,amino, or mercapto, are or need to be protected in a compound offormulae I, because they should not take part in the reaction, these aresuch groups as are usually used in the synthesis of peptide compounds,and also of cephalosporins and penicillins, as well as nucleic acidderivatives and sugars.

The protecting groups may already be present in precursors and shouldprotect the functional groups concerned against unwanted secondaryreactions, such as acylations, etherifications, esterifications,oxidations, solvolysis, and similar reactions. It is a characteristic ofprotecting groups that they lend themselves readily, i.e. withoutundesired secondary reactions, to removal, typically by solvolysis,reduction, photolysis or also by enzyme activity, for example underconditions analogous to physiological conditions, and that they are notpresent in the end-products. The specialist knows, or can easilyestablish, which protecting groups are suitable with the reactionsmentioned hereinabove and hereinafter.

The starting compounds of formula II and formula III may be produced asdescribed in the Examples.

The compounds of the invention exhibit valuable pharmacologicalproperties in mammals and are therefore useful as pharmaceuticals. Theyare particularly useful as inhibitors of cathepsin K or cathepsin S orboth.

The cathepsin K inhibitory effects of the compound of the invention canbe demonstrated in vitro by measuring the inhibition of e.g. recombinanthuman cathepsin K.

The in vitro assay is carried out as follows:

The assay is performed in 96 well microtiter plates at ambienttemperature using recombinant human cathepsin K. Inhibition of cathepsinK is assayed at a constant enzyme (0.16 nM) and substrate concentration(54 mm Z-Phe-Arg-AMCA—Peptide Institute Inc. Osaka, Japan) in 100 mMsodium phosphate buffer, pH 7.0, containing 2 mM dithiothreitol, 20 mmTween 80 and 1 mm EDTA. Cathepsin K is preincubated with the inhibitorsfor 30 min and the reaction is initiated by the addition of substrate.After 30 min incubation the reaction is stopped by the addition of E-64(2 mm), and fluorescence intensity is read on a multi-well plate readerat excitation and emission wavelengths of 360 and 460 nm, respectively.

Compounds of the Invention typically have IC₅₀s for inhibition of humancathepsin K of less than about 100 nM down to about 1 nM or less,preferably of about 5 nM or less, e.g. about 0.2 nM. Example 3-0 has aIC₅₀ in the above described assay of about 0.16 nM. Preferred arecompounds as defined above with R=R1, e.g. compounds of examples 1 to 4with R=R1, which have cathepsin K inhibitory effects. More preferred arecompounds as defined above with R=R5, most preferred Example 3-0.

The cathepsin S inhibitory effects of the compound of the invention canbe demonstrated in vitro by measuring the inhibition of e.g. recombinanthuman cathepsin S.

The in vitro assay is carried out in clear, flat-bottomed, 96-wellmicrotiter plates (Greiner GmbH, Germany) at ambient temperature usingrecombinant human cathepsin S. Inhibition of human cathepsin S isassayed at a constant enzyme and various substrate concentrations(substrate is Z-Leu-Leu-4-methylcoumaryl-7-amide (Bachem (Switzerland))in 100 parts 0.2M sodium phosphate, pH 7.0, containing 2 mM EDTA, 2parts 1% Triton X-100, 10 parts 20 mM dithiothreitol (DTT) and 58 partsdistilled water. The assay is started by adding the enzyme solution (13times higher concentration of final concentration of recombinant humanCathepsin S) to the reaction mixture containing various concentrationsof the corresponding substrate and the compound. Substrateconcentrations between 3.4 and 17 μM are used. The recombinant humanCathepsin S is used at a final concentration of 0.04 nM. Test compoundsare used at concentrations between 0.4 and 2 times the determined IC₅₀of the compound at the enzyme. The relative fluorescence is continuouslymeasured for 30 minutes and the initial velocity is obtained from eachprogress curve. The inhibition patterns and the K_(i) values aredetermined by Dixon plot analysis.

Compounds of the Invention typically have IC₅₀s for inhibition of humancathepsin S of less than about 100 nM down to about 1 nM or less,preferably of about 5 nM or less. Preferred compounds are compounds asdefined above with R=R2.E.g. example 4-8 has a IC₅₀ in the abovedescribed assay of about 9 nM.

Compounds of the Invention which have dual inhibitory effects, i.e.inhibitory effects in the cathepsin K and the cathepsin S assay asdescribed above typically have IC₅₀s for inhibition of human cathepsin Sand of cathepsin K of less than about 100 nM in both assays, down toabout 1 nM or less in both assays, preferably of about 5 nM or less.Preferred compounds with a dual inhibitory effect are compounds asdefined above with R=R6. E.g. example 4-3 with an IC₅₀ on humancathepsin K of 8 nM and on human cathepsin S of 6 nM. Or example 4-9with an IC₅₀ on human cathepsin K of 16 nM and on human cathepsin S of10 nM.

In view of their activity as inhibitors of cathepsin K and/or cathepsinS, compounds of the invention are particularly useful in mammals asagents for treatment and prophylaxis of diseases and medical conditionsinvolving elevated levels of cathepsin K and or cathepsin S. Suchdiseases include diseases involving infection by organisms such aspneumocystis carinii, trypsanoma cruzi, trypsanoma brucei, crithidiafusiculata, as well as parasitic diseases such as schistosomiasis andmalaria, tumours (tumour invasion and tumour metastasis), and otherdiseases such as metachromatic leukodystrophy, muscular dystrophy,amytrophy, neuropathic pain, e.g. chronic neuropathic pain, exemplifiedby conditions such as diabetic neuropathy, postherpetic neuralgia,trigeminal neuralgia, painful diabetic polyneuropathy, post-stroke pain(central pain), postamputation pain, myolopathic or radiculopathic pain(e.g. spinal stenosis, arachnoiditis, root sleeve fibrosis), atypicalfacial pain and causalgia-like syndromes (complex regional painsyndromes), autoimmune disorders, including, but not limited to juvenileonset diabetes and multiple sclerosis, allergic disorders, including,but not limited to, asthma, and allogeneic immune responses, including,but not limited to, organ transplant rejection.

In particular, cathepsin K has been implicated in diseases of excessivebone loss, and thus the Compounds of the Invention may be used fortreatment and prophylaxis of such diseases, including osteoporosis,osteoporosis of various genesis (e.g. juvenile, menopausal,post-menopausal, post-traumatic, caused by old age or by cortico-steroidtherapy or inactivity), gingival diseases such as gingivitis andperiodontitis, Paget's disease, hypercalcemia of malignancy, e.g.tumour-induced hypercalcemia and metabolic bone disease. Also theCompounds of the Invention may be use for treatment or prophylaxis ofdiseases of excessive cartilage or matrix degradation, includingosteoarthritis and rheumatoid arthritis as well as certain neoplasticdiseases involving expression of high levels of proteolytic enzymes andmatrix degradation. Preferably cathepsin K inhibitors are used in thetreatment of osteoporosis and osteoarthritis.

In particular, cathepsin S has been implicated in the treatment and alsoin the prevention of neuropathic pain, e.g. chronic neuropathic pain,exemplified by conditions such as diabetic neuropathy, postherpeticneuralgia, trigeminal neuralgia, painful diabetic polyneuropathy,post-stroke pain (central pain), postamputation pain, myolopathic orradiculopathic pain (e.g. spinal stenosis, arachnoiditis, root sleevefibrosis), atypical facial pain and causalgia-like syndromes (complexregional pain syndromes), osteoarthritis and rheumatoid arthritis,autoimmune disorders, including, but not limited to juvenile onsetdiabetes and multiple sclerosis, allergic disorders, including, but notlimited to, asthma, and allogeneic immune responses, including, but notlimited to, organ transplant rejection. Preferably cathepsin Sinhibitors are used in the treatment of neuropathic pain, multiplesclerosis, osteoarthritis and rheumatoid arthritis.

Dual inhibitors may thus be implicated in diseases of where bothcathepsins play a role, e.g. neuropathic pain, inflammation, rheumatoidarthritis, osteoarthritis, osteoporosis, tumors (especially tumorinvasion and tumor metastasis), obesity, coronary disease,atherosclerosis (including atherosclerotic plaque rupture anddestabilization), autoimmune diseases, multiple sclerosis, respiratorydiseases, infectious diseases and immunologically mediated diseases(including transplant rejection), preferably neuropathic pain,osteoporosis, rheumatoid arthritis, and osteoarthritis.

Beneficial effects are evaluated in in vitro and in vivo pharmacologicaltests generally known in the art, and as illustrated herein. The abovecited properties are demonstrable in in vitro and in vivo tests, usingadvantageously mammals, e.g. rats, mice, dogs, rabbits, monkeys orisolated organs and tissues, as well as mammalian enzyme preparations,either natural or prepared by e.g. recombinant technology. Compounds ofthe Invention can be applied in vitro in the form of solutions, e.g.preferably aqueous solutions or suspensions, and in vivo eitherenterally or parenterally, advantageously orally, e.g. as a suspensionor in aqueous solution, or as a solid capsule or tablet formulation. Thedosage in vitro may range between about 10⁻⁵ molar and 10⁻⁹ molarconcentrations. The dosage in vivo may range, depending on the route ofadministration, between about 0.1 and 100 mg kg.

The efficacy of the Compounds of the Invention for the treatment ofosteoporosis can be determined using the In vivo animal model “OVXcynomolgus monkey”. This model is well known in the art and is a commonmodel to validate an osteoporosis compound (see e.g. Jerome C P,Peterson P E (2001) Bone; 29 (1): 1-6).

The efficacy of the Compounds of the Invention for the treatment ofchronic inflammatory or neuropathic pain can be determined using thefollowing In vivo animal models:

Chronic Inflammatory Pain Model:

The Complete Freund's Adjuvant—induced mechanical hyperalgesia may beused as a model of chronic inflammatory pain (Stein, C. et al.Pharmacol. Biochem. Behav. (1988) 31:445-451). In this model, typicallya male Sprague-Dawley or Wistar rat (200-250 g) receives an intraplantarinjection of 25 μl complete Freund's adjuvant into one hind paw. Amarked inflammation occurs in this hind paw. Drugs are generallyadministered for evaluation of efficacy, 24 hours after the inflammatoryinsult, when mechanical hyperalgesia is considered fully established.

Chronic Neuropathic Pain Models:

Two animal models of chronic neuropathic pain may be used that involvesome form of peripheral nerve damage. In the Seltzer model (Seltzer etal. (1990) Pain 43: 205-218) rats are anaesthetised and a small incisionmade mid-way up one thigh (usually the left) to expose the sciaticnerve. The nerve is carefully cleared of surrounding connective tissuesat a site near the trochanter just distal to the point at which theposterior biceps semitendinosus nerve branches off the common sciaticnerve. A 7-0 silk suture is inserted into the nerve with a ⅜ curved,reversed-cutting mini-needle, and tightly ligated so that the dorsal ⅓to ½ of the nerve thickness is held within the ligature. The muscle andskin are closed with sutures and clips and the wound dusted withantibiotic powder. In sham animals the sciatic nerve is exposed but notligated and the wound closed as in nonsham animals.

In the Chronic Constriction Injury (CCI) model (Bennett, G. J. and Xie,Y. K. Pain (1988) 33: 87-107) rats are anaesthetised and a smallincision is made mid-way up one thigh (usually the left) to expose thesciatic nerve. The nerve is cleared of surrounding connective tissue andfour ligatures of 4/0 chromic gut are tied loosely around the nerve withapproximately 1 mm between each, so that the ligatures just barelyconstrict the surface of the nerve. The wound is closed with sutures andclips as described above. In sham animals the sciatic nerve is exposedbut not ligated and the wound closed as in nonsham animals.

In contrast to the Seltzer and CCI models, the Chung model involvesligation of the spinal nerve. (Kim, S. O. and Chung, J. M. Pain (1992):50:355-363). In this model, rats are anesthetized and placed into aprone position and an incision is made to the left of the spine at theL4-S2 level. A deep dissection through the paraspinal muscles andseparation of the muscles from the spinal processes at the L4-S2 levelwill reveal part of the sciatic nerve as it branches to form the L4, L5and L6 spinal nerves. The L6 transverse process is carefully removedwith a small rongeur enabling visualisation of these spinal nerves. TheL5 spinal nerve is isolated and tightly ligated with 7-0 silk suture.The wound is closed with a single muscle suture (6-0 silk) and one ortwo skin closure clips and dusted with antibiotic powder. In shamanimals the L5 nerve is exposed as before but not ligated and the woundclosed as before.

Behavioral Index

In all chronic pain models (inflammatory and neuropathic) mechanicalhyperalgesia is assessed by measuring paw withdrawal thresholds of bothhindpaws to an increasing pressure stimulus using an Analgesymeter(Ugo-Basile, Milan). Mechanical allodynia is assessed by measuringwithdrawal thresholds to non-noxious mechanical stimuli applied with vonFrey hairs to the plantar surface of both hindpaws. Thermal hyperalgesiais assessed by measuring withdrawal latencies to a noxious thermalstimulus applied to the underside of each hindpaw. With all models,mechanical hyperalgesia and allodynia and thermal hyperalgesia developwithin 1-3 days following surgery and persist for at least 50 days. Forthe assays described herein, drugs may be applied before and aftersurgery to assess their effect on the development of hyperalgesia,particularly approximately 14 days following surgery, to determine theirability to reverse established hyperalgesia.

The percentage reversal of hyperalgesia is calculated as follows:

${\% \mspace{14mu} {reversal}} = {\frac{{{postdose}\mspace{14mu} {threshold}} - {{predose}\mspace{14mu} {threshold}}}{{{naive}\mspace{14mu} {threshold}} - {{predose}\mspace{14mu} {threshold}}} \times 100}$

In the experiments disclosed herein, Wistar rats (male) are employed inthe pain models described above. Rats weigh approximately 120-140 gramsat the time of surgery. All surgery is performed under enflurane/O₂inhalation anaesthesia. In all cases the wound is closed after theprocedure and the animal allowed to recover. In all pain modelsemployed, after a few days in all but the sham operated animals, amarked mechanical and thermal hyperalgesia and allodynia develops inwhich there is a lowering of pain threshold and an enhanced reflexwithdrawal response of the hind-paw to touch, pressure or thermalstimuli. After surgery the animals also exhibit characteristic changesto the affected paw. In the majority of animals the toes of the affectedhind paw are held together and the foot turned slightly to one side; insome rats the toes are also curled under. The gait of the ligated ratsvaries, but limping is uncommon. Some rats are seen to raise theaffected hind paw from the cage floor and to demonstrate an unusualrigid extension of the hind limb when held. The rats tend to be verysensitive to touch and may vocalise. Otherwise the general health ancondition of the rats is good.

Compounds of the invention, are also indicated for preventing ortreating coronary disease, atherosclerosis (including atheroscleroticplaque rupture and destabilization) (see e.g. Cathepsin F and S blockHDL3-induced cholesterol efflux from macrophage cells, Lindstedt et al.,2003, Biochemical and Biophysical research communications 312:1019-1024), autoimmune diseases, respiratory diseases andimmunologically mediated diseases (including transplant rejection).

The antiarthritic efficacy of the compounds of the invention for thetreatment of rheumatoid arthritis can be determined using models such asor similar to the rat model of adjuvant arthritis, as describedpreviously (R. E. Esser, et. al. J. Rheumatology, 1993, 20, 1176.)

The efficacy of the compounds of the invention for the treatment ofosteoarthritis can be determined using models such as or similar to therabbit partial lateral meniscectomy model, as described previously(Colombo et al. Arth. Rheum. 1993 26, 875-886). The efficacy of thecompounds in the model can be quantified using histological scoringmethods, as described previously (O'Byrne et al. Inflamm Res 1995,S117-S118).

The efficacy of the compounds of the invention for the treatment ofosteoporosis can be determined using an animal model such as theovariectomised rat or other similar species, e.g. rabbit or monkey, inwhich test compounds are administered to the animal and the presence ofmarkers of bone resorption are measured in urine or serum (e.g. asdescribed in Osteoporos Int (1997) 7:539-543).

Accordingly in further aspects the invention provides:

A compound of the invention for use as a pharmaceutical; apharmaceutical composition comprising a compound of the invention as anactive ingredient; a method of treating a patient suffering from orsusceptible to a disease or medical condition in which cathepsin Kand/or cathepsin S is implicated, comprising administering an effectiveamount of a compound of the invention to the patient, and the use of acompound of the invention for the preparation of a medicament fortherapeutic or prophylactic treatment of a disease or medical conditionin which cathepsin K and or cathepsin S is implicated.

The present invention relates to methods of using compounds of theinvention and their pharmaceutically acceptable salts, or pharmaceuticalcompositions thereof, in mammals for inhibiting cathepsin K and orcathepsin S, and for the treatment of cathepsin K and or cathepsin Sdependent conditions, such as the cathepsin K and or cathepsin Sdependent conditions, described herein, e.g. inflammation, neuropathicpain, osteoporosis, rheumatoid arthritis and osteoarthritis.

In a particular embodiment of the invention, the present inventionrelates to a method of selectively inhibiting cathepsin K activity in amammal which comprises administering to a mammal in need thereof aneffective cathepsin K inhibiting amount of a compound of the invention.

More specifically such relates to a method of treating osteoporosis,rheumatoid arthritis, osteoarthritis, and inflammation (and otherdiseases as identified above) in mammals comprises administering to amammal in need thereof a correspondingly effective amount of a compoundof the invention.

In a particular embodiment of the invention, the present inventionrelates to a method of selectively inhibiting cathepsin S activity in amammal which comprises administering to a mammal in need thereof aneffective cathepsin S inhibiting amount of a compound of the invention.

More specifically such relates to a method of treating neuropathic pain(and other diseases as identified above) in mammals comprisesadministering to a mammal in need thereof a correspondingly effectiveamount of a compound of the invention.

In a particular embodiment of the invention, the present inventionrelates to a method of inhibiting cathepsin K and cathepsin S activityin a mammal which comprises administering to a mammal in need thereof aneffective cathepsin K and cathepsin S inhibiting amount of a compound ofthe invention.

More specifically such relates to a method of treating neuropathic pain,osteoporosis, rheumatoid arthritis, osteoarthritis, and inflammation(and other diseases as identified above) in mammals comprisesadministering to a mammal in need thereof a correspondingly effectiveamount of a compound of the invention.

The following examples are intended to illustrate the invention and arenot to be construed as being limitations thereon. Temperatures are givenin degrees Centigrade. If not mentioned otherwise, all evaporations areperformed under reduced pressure, preferably between about 15 and 100 mmHg (=20-133 mbar). The structure of final products, intermediates andstarting materials is confirmed by standard analytical methods, e.g.microanalysis and spectroscopic characteristics (e.g. MS, IR, NW)Abbreviations used are those conventional in the art.

EXAMPLES Example 1 Example 1-0 Preparation of7-(2,2-Dimethyl-propyl)-6-[2-(2-hydroxy-ethyl)-1,3-dioxo-2,8-diaza-spiro[4.5]dec-8-ylmethyl]-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

A. Preparation of 8-Benzyl-2,8-diaza-spiro[4.5]decane-1,3-dione

To a solution of 1-benzyl-piperidin-4-one (75.1 g, 0.40 mol) in toluene(400 ml), cyano-acetic acid ethyl ester (50.6 ml, 0.48 mol) and aceticacid (18.2 ml, 0.32 mol) are added at ambient temperature. The reactionmixture is refluxed for 4 h, quenched with ice-water and extracted withdiethyl ether. The combined extracts are washed with H₂O, brine anddried over sodium sulphate to give(1-benzyl-piperidin-4-ylidene)-cyano-acetic acid ethyl ester inquantitative yield.

Rf=0.53 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.30-1.37 (m, 3H), 2.58 (dd, 2H), 2.64 (dd,2H), 2.79 (dd, 2H), 3.15 (dd, 2H), 3.55 (s, 2H), 4.23-4.32 (m, 2H),7.21-7.36 (m, 5H).

To a solution of (1-benzyl-piperidin-4-ylidene)-cyano-acetic acid ethylester (112.9 g, 0.40 mol) in EtOH (500 ml) and H₂O (100 ml), potassiumcyanide (64.6 g, 0.99 mol) is added at ambient temperature. The reactionmixture is stirred at 65 C.° for 24 h. After removal of EtOH, H₂O isadded to the residue. The waster phase is extracted with diethyl ether.The combined extracts are washed with H₂O and brine, dried over sodiumsulfate and evaporated down to give 77.7 g of1-benzyl-4-cyanomethyl-piperidine-4-carbonitrile.

Rf=0.38 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz CDCl₃) δ: 1.76-1.81 (m, 2H), 2.10-2.05 (m, 2H),2.23-2.39 (m, 2H), 2.69 (s, 2H), 2.90-2.94 (m, 2H), 3.56 (s, 2H),7.21-7.38 (m, 5H).

Acetic acid (56.8 ml) and sulfuric acid (11.8 ml) are added to1-benzyl-4-cyanomethyl-piperidine-4-carbonitrile (27.2 g, 0.114=mol) atambient temperature. The reaction mixture is stirred at 125 C.° for 1 h,cooled down to the room temperature and added to saturated NaOH aq. toadjust to pH 6.0. The mixture is extracted with dichloromethane. Thecombined extracts are washed with H₂O and brine, dried over sodiumsulfate and evaporated down to give8-benzyl-2,8-diaza-spiro[4.5]decane-1,3-dione (three steps yield:81.8%).

Rf=0.40 (CH₂Cl₂:MeOH=10:1).

¹H-NMR (400 MHz CDCl₃) δ: 1.52-1.57 (m, 2H), 2.02-2.17 (m, 4H), 2.59 (s,2H), 2.86-2.90 (m, 2H), 3.52 (s, 2H), 7.21-7.28 (m, 2H), 7.30-7.37 (m,3H), 7.92 (brs, 1H).

B. Preparation of 1,3-Dioxo-2,8-diaza-spiro[4.5]decane-8-carboxylic acidtert-butyl ester

To 8-benzyl-2,8-diaza-spiro[4.5]decane-1,3-dione (28.3 g, 0.11 mol) andPd(OH)₂ (8.5 g) in 2 l of flask, EtOH (438 ml) and acetic acid (5.5 ml)are added at ambient temperature. The reaction mixture is stirred underH₂ at room temperature for 15 h. The catalysts are removed by filtrationand EtOH is evaporated down to give 2,8-diaza-spiro[4.5]decane-1,3-dionein quantitative yield. To a suspension of2,8-diaza-spiro[4.5]decane-1,3-dione (4.2 g, 25.2 mmol) indichloromethane (60 ml), 1N NaOH (26 ml, 26 mmol) anddi-t-butyldicarbonate (6.1 g, 27.7 mmol) in dichloromethane (20 ml) areadded at ambient temperature. The reaction mixture is stirred for 15 h.10% Citric acid is added to the reaction mixture and the pH of themixture is adjusted to 5. The combined extracts are washed with brine,dried over magnesium sulfate and concentrated under vacuum to give solidproduct, which filtrated with diethyl ether.

Yield: 51%

Rf=0.25 (n-hexane:ethyl acetate=1:1).

¹H-NMR (400 M Z CDCl₃) δ: 1.47 (s, 9H), 1.55-1.70 (m, 2H), 1.95-2.05 (m,2H), 2.62 (s, 2H), 2.96-3.02 (m, 2H), 4.02-4.04 (m, 2H), 8.14 (brs, 1H).

Preparation of1,3-dioxo-2-[2-(tetrahydro-pyran-2-yloxy)-ethyl]-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester

To a suspension of 1,3-dioxo-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester (1.0 g, 3.7=mol) in DMF (12 ml),2-(2-bromoethoxy)-tetrahydro-2H-pyrane (0.62 ml, 4.1 mmol) and potassiumcarbonate (0.62 g, 4.5 mmol) are added at ambient temperature and themixture is stirred for overnight at room temperature. The reactionmixture is quenched with water and extracted with ethyl acetate. Thecombined extracts are washed with brine and dried over sodium sulfate,filtrated. The solvent is evaporated down to give 1.6 g of crude1,3-dioxo-2-[2-(tetrahydro-pyran-2-yloxy)-ethyl]-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester.

Preparation of 2-(2-hydroxy-ethyl)-2,8-diaza-spiro[4.5]decane-1,3-dione.hydrochloride

To a solution of crude1,3-dioxo-2-[2-(tetrahydro-pyran-2-yloxy)-ethyl]-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester (1.6 g) in ethyl acetate (1 ml), EtOH (1.0 ml) and4N HCl/ethyl acetate (4 ml) are added at room temperature. The reactionmixture is stirred for overnight at room temperature. The solvent isremoved by evaporation to give 1.06 g of crude2-(2-hydroxy-ethyl)-2,8-diaza-spiro[4.5]decane-1,3-dione. hydrochloride.

Preparation of7-(2,2-Dimethyl-propyl)-6-[2-(2-hydroxy-ethyl)-1,3-dioxo-2,8-diaza-spiro[4.5]dec-8-ylmethyl]-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

6-Bromomethyl-7-(2,2-dimethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile(1.3 g, 4.25 mmol) and crude2-(2-hydroxy-ethyl)-2,8-diaza-spiro[4.5]decane-1,3-dione. hydrochloride(1.1 g, 4.25 mmol) are dissolved in DMF (14 ml) and potassium carbonate(1.8 g, 12.8 mmol) is added to the solution. The reaction mixture isstirred at room temperature for 12 h and quenched with H₂O and extractedwith ethyl acetate. The combined extracts are washed with brine, driedover magnesium sulfate and evaporated down. The crude product ispurified by reverse phase HPLC and fraction are collected and evaporateddown. Saturated sodium bicarbonate is added and neutralized and thewater phase is extracted with ethyl acetate. The combined extracts arewashed with brine, dried over magnesium sulfate and evaporated down togive 0.5 g of desired product in 27% yield.

Rf=0.10 (n-hexane:ethyl acetate=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.01 (s, 9H), 1.52-1.60 (m, 2H), 2.08-2.14(m, 4H), 2.60 (s, 2H), 2.84-2.88 (m, 2H), 3.71-3.78 (m, 4H), 3.81 (s,2H), 4.34 (s, 2H), 6.58 (s, 1H), 8.89 (s, 1H).

By repeating the procedures described above using appropriate startingmaterials and conditions the following compounds of formula I areobtained as identified below in Table 1.

TABLE 1 Formula I-(i)

Yield Example Rx (%) Rf (Solvent) NMR (400 MHz, δ) I-1 H 43 0.48(CDCl₃): 1.03 (s, 9H), 1.50-1.57 (EtOAc only) (m, 2H), 2.10-2.25 (m,4H), 2.62 (s, 2H), 2.84-2.95 (m, 2H), 3.83 (s, 2H), 4.36 (s, 2H), 6.60(s, 1H), 7.74 (brs, 1H), 8.91 (s, 1H) 1-2

40 0.20 (n-hexane:AcOEt = 1:1) (CDCl₃): 1.01 (s, 9H), 1.48-1.52 (m, 2H),2.11-2.16 (m, 4H), 2.55 (s, 2H), 2.84-2.87 (m, 2H), 3.00 (s, 3H), 3.81(s, 2H), 4.35 (s, 2H), 6.58 (s, 1H), 8.89 (s, 1H) 1-3

30 0.50 (n-hexane:AcOEt = 1:1) (CDCl₃): ): 0.88 (t, 3H), 1.01 (s, 9H),1.51-1.65 (m, 4H), 2.07- 2.19 (m, 4H), 2.54 (s, 2H), 2.84-2.87 (m, 2H),3.44-3.48 (m, 2H), 3.81 (s, 2H), 4.35 (s, 2H), 6.58 (s, 1H), 8.89 (s,1H) 1-4

35 0.50 (n-hexane:AcOEt = 1:1) (CDCl₃): 1.02 (s, 9H), 1.37 (d, 6H),1.47-1.50 (m, 2H), 2.04- 2.20 (m, 4H), 2.51 (s, 2H), 2.84-2.87 (m, 2H),3.82 (s, 2H), 4.35-4.37 (m, 3H), 6.61 (s, 1H), 8.89 (s, 1H) 1-5

42 0.20 (n-hexane:AcOEt = 1:1) (CDCl₃): 1.01 (s, 9H), 1.51- 1.54 (m,2H), 2.07-2.17 (m, 4H), 2.40-2.50 (m, 4H), 2.52- 2.55(m, 4H), 2.83-2.86(m, 2H), 3.57-3.61 (m, 6H), 3.81 (s, 2H), 4.35 (s, 2H), 6.58 (s, 1H),8.89 (s, 1H) 1-6

50 0.40 (n-hexane:AcOEt = 1:1) (CDCl3): 1.00 (s, 9H), 1.46- 1.49 (m,2H), 2.04-2.14 (m, 4H), 2.55 (s, 2H), 2.81-2.84 (m, 2H), 3.79 (s, 2H),4.34 (s, 2H), 4.60 (s, 2H), 6.57 (s, 1H), 6.95-6.99 (m, 2H), 7.31-7.35(m, 2H), 8.88 (s, 1H) 1-7

26 0.250 (n-hexane:AcOEt = 1:1) (CDCl3): 1.00 (s, 9H), 1.48- 1.52 (m,2H), 2.06-2.16 (m, 4H), 2.57 (s, 2H), 2.82-2.86 (m, 2H), 3.80 (s, 2H),4.34 (s, 2H), 4.68 (s, 2H), 6.57 (s, 1H), 6.75-6.83 (m, 2H), 7.26-7.30(m, 1H), 8.89 (s, 1H) 1-8

19 0.40 (n-hexane:AcOEt = 1:1) (CDCl3): 1.00 (s, 9H), 1.47- 1.51 (m,2H), 2.07-2.18 (m, 4H), 2.54 (s, 2H), 2.82-2.85 (m, 2H), 3.76 (s, 6H),3.80 (s, 2H), 4.35 (s, 2H), 4.62 (s, 2H), 6.38-6..40 (m, 2H), 6.58 (s,1H), 7.06 (d, 1H), 8.88 (s, 1H) 1-9

23 0.40 (n-hexane:AcOEt = 1:1) (CDCl3): 1.00 (s, 9H), 1.45- 1.49 (m,2H), 2.06-2.14 (m, 4H), 2.54 (s, 2H), 2.81-2.84 (m, 2H), 3.79 (s, 2H),3.84 (s, 6H), 4.34 (s, 2H), 4.57 (s, 2H), 6.57 (s, 1H), 6.77 (d, 1H),6.91- 6.93 (m, 2H), 8.88 (s, 1H)

Example 2 Example 2-0 Preparation7-(2,2-Dimethyl-propyl)-6-(5methoxy-2-oxo-1,2-dihydro-1′H-spiro[indole-3,4′-piperdine]-1-ylmethyl)-7-pyrrole[2,3d]pyrimidine-2-carbonitrile

A. Preparation of 2-Fluoro-4-methoxy-1-nitro-benzene

To a solution of 3-fluoro-4-nitro-phenol (25.3 g, 0.16 mol) in acetone(160 ml), potassium carbonate (41.7 g, 0.30 mol) and methyl iodide (20.0ml, 0.32 mol) are added at ambient temperature. The reaction mixture isstirred at 40 C.° for 3 h. After cooling down to room temperature,dichloromethane is added to the reaction mixture, which is filtrated andevaporated. Dichloromethane is added to the residue and the organicphase is washed with H₂O and brine, dried over sodium sulfate andevaporated down to give 2-fluoro-4-methoxy-1-nitro-benzene in 98% yield.

Rf=0.5 (n-hexane:ethyl acetate=10:1).

¹H-NMR (400 MHz, CDCl₃) δ: 3.90 (s, 3H), 6.72-6.79 (m, 2H), 8.06-8.13(m, 1H).

B. Preparation of 5-Methoxy-1,3-dihydro-indol-2-one

The title compound was prepared according to method reported in thepatent (WO0206228).

To a solution of 2-fluoro-4-methoxy-1-nitro-benzene (84.1 g, 0.49 mol)and dimethyl malonate (129.9 g, 0.98 mol) in DMF (490 ml), potassiumcarbonate (135.9 g, 0.98 mol) is added at ambient temperature. Thereaction mixture is stirred at 70 C.° for 12 h. The reaction mixture isadded to toluene (393 ml) and 12 N HCl (123 ml) and extracted with ethylacetate. The combined extracts are washed with H₂O and brine, dried oversodium sulfate and evaporated down to give2-(5-methoxy-2-nitro-phenyl)-malonic acid dimethyl ester.

Rf=0.8 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 3.80 (s, 6H), 3.89 (s, 3H), 5.45 (s, 1H),6.94-6.96 (m, 2H), 8.15-8.20 (m, 1H).

To 2-(5-methoxy-2-nitro-phenyl)-malonic acid dimethyl ester and 5% Pd—C(7.0 g) in 1 l of flask, MeOH (490 ml) is added at ambient temperature.The reaction mixture is stirred under H₂ at room temperature for 15 h.The catalysts are removed by filtration and MeOH is evaporated down togive crude 5-methoxy-2-oxo-2,3-dihydro-1H-indole-3-carboxylic acidmethyl ester.

Rf=0.10 (n-hexane:ethyl acetate=1:1).

To a solution of crude5-methoxy-2-oxo-2,3-dihydro-1H-indole-3-carboxylic acid methyl ester inMeOH (320 ml), 6N HCl (255 ml, 1.92 mol) is added at ambienttemperature. The reaction mixture is stirred at 70 C.° for 3 h. Aftercooling down to room temperature, 8 N KOH (269 ml, 1.82 mol) is added toreaction mixture. The reaction mixture is stirred at 40 C.° for 30 min.12 N HCl (41 ml) is added to reaction mixture. MeOH is evaporated downand the white powder is filtrated.

Yield: 59% (three steps).

Rf=0.25 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 3.51 (s, 2H), 3.78 (s, 3H), 6.72-6.85 (m,3H), 7.60 (brs, 1H).

C. Preparation of1′-Benzyl-5-methoxyspiro[indole-3,4′-piperidin]-2(1H)-one

To a solution of NaHMDS (1 M THF solution) (800 ml, 0.8 mol), thesolution of 5-methoxy-1,3-dihydro-indol-2-one (26.1 g, 0.16 mol) in THF(160 ml) and benzyl-bis-(2-chloro-ethyl)-amine (47.3 g, 0.18 mol) in THF(176 ml) are added at −78° C. The reaction mixture is stirred for 15 hat room temperature, quenched with saturated ammonium chloride andice-water and extracted with ethyl acetate. The combined extracts arewashed with brine, dried over sodium sulphate and evaporated down. Ethylether is added to the residue to give the powder, which is filtrated.

Yield: 39%

Rf=0.25 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.81-1.99 (m, 2H), 2.00-2.04 (m, 2H),2.66-2.72 (m, 2H), 2.90-2.96 (m, 2H), 3.67 (s, 2H), 3.77 (s, 3H),6.71-6.81 (m, 2H), 7.00 (s, 1H), 7.25-7.40 (m, 5H), 8.32 (brs, 1H).

D. Preparation of tert-Butyl5-methoxy-2-oxo-1,2-dihydro-1′H-spiro[indole-3,4′-piperidine]-1′-carboxylate

To 1′-benzyl-5-methoxyspiro[indole-3,4′-piperidin]-2(1H)-one (20.0 g, 62mmol) and Pd C (2.0 g) in 500 ml of flask, EtOH (120 ml) and acetic acid(5.5 ml) are added at ambient temperature. The reaction mixture isstirred under H₂ at room temperature for 15 h. The catalysts are removedby filtration and EtOH is evaporated down.

Rf=0.20 (n-hexane:AcOEt=1:1).

To a suspension of 5-methoxyspiro[indole-3,4′-piperidin]-2(1H)-one (9.9g, 45.2 mmol) in dichloromethane (50 ml), 1N NaOH (45.2 ml, 45.2 mmol)and the solution of di-t-butyldicarbonate (9.3 g, 45.2 mmol) indichloromethane (50 ml) are added at ambient temperature. The reactionmixture is stirred for 1 h. The reaction mixture is washed with brine,dried over magnesium sulfate and concentrated under vacuum.Chromatography on silica gel (eluent; n-hexane:ethyl acetate=2:1 and1:1) gives 10.6 g of desired product.

Yield: 68% (two steps)

Rf=0.50 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.51 (s, 9H), 1.76-1.89 (m, 4H), 3.70-3.90(m, 7H), 6.74-6.76 (m, 1H), 6.83-6.88 (m, 2H), 8.83 (brs, 1H).

E. Preparation of1-[2-Cyano-7-(2,2-dimethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-5-methoxy-2-oxo-1,2-dihydro-1′H-spiro[indole-3,4′-piperidine]-1′carboxyric acid tert-butyl ester

To a solution of tert-butyl5-methoxy-2-oxo-1,2-dihydro-1′H-spiro[indole-3,4′-piperidine]-1′-carboxylate(10.6 g, 31.9 mmol) in DMF (70 ml), NaH (1.4 g, 35.1 mmol) are added atroom temperature and the mixture is stirred at room temperature for 30min.6-Bromomethyl-7-(2,2-dimethyl-propyl)-7.H.-pyrrolo[2,3-.d.]pyrimidine-2-carbonitrile(9.5 g, 31.9 mmol) is added at 0° C. and the reaction mixture is stirredfor 4 h at ambient temperature. The reaction mixture is quenched withice-water and extracted with ethyl acetate. The combined extracts arewashed with brine and dried over magnesium sulfate. Chromatography onsilica gel (eluent; n-hexane:ethyl acetate=10:1 and 1:1) give 12.1 g oftitle product.

Yield: 68%

Rf=0.60 (n-hexane:ethyl acetate=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.09 (s, 9H), 1.50 (s, 9H), 1.83-1.86 (m,4H), 3.78-3.84 (m, 7H), 4.22 (s, 2H), 5.13 (s, 2H), 6.37 (s, 1H),6.62-6.65 (m, 1H), 6.72-6.75 (m, 1H), 7.26 (s, 1H), 8.84 (s, 1H).

F. Preparation of7-(2,2-Dimethyl-propyl)-6-(5-methoxy-2-oxo-1,2-dihydro-1′H-spiro[indole-3,4′-piperidine]-1-ylmethyl)-7-pyrrole[2,3d]pyrimidine-2-carbonitrile

To a solution of1-[2-cyano-7-(2,2-dimethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-5-methoxy-2-oxo-1,2-dihydro-1′H-spiro[indole-3,4′-piperidine]-1′carboxyric acid tert-butyl ester (12.1 g, 21.6 mmol) in dichloromethane(100 ml), TFA (5 ml) is added at 0° C. The reaction mixture is stirredat room temperature for 2 h. After removal of the solvent, saturatedsodium bicarbonate is added to the residue and the mixture is extractedwith ethyl acetate. The combined extracts are washed with brine, driedover magnesium sulfate and concentrated under vacuum. Ethyl ether isadded to the residue, which is filtrated to give pale yellow product,7-(2,2-Dimethyl-propyl)-6-(5methoxy-2-oxo-1,2-dihydro-1′H-spiro[indole-3,4′-piperidine]-1-ylmethyl)-7-pyrrole[2,3d]pyrimidine-2-carbonitrile.

Yield: 91%.

Rf=0.15 (CH₂Cl₂:MeOH=10:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.09 (s, 9H), 1.90-1.94 (m, 2H), 2.52-2.61(m, 2H), 3.4-3.50 (m, 2H), 3.80 (s, 3H), 3.87-3.94 (m, 2H), 4.24 (s,2H), 5.12 (s, 2H), 6.37 (s, 1H), 6.67 (d, 1H) 6.77-6.80 (m, 1H), 7.03(s, 1H), 8.86 (s, 1H).

By repeating the procedures described above using appropriate startingmaterials and conditions the following compounds of formula I-(ii) areobtained as identified below in Table 2:

TABLE 2 Formula I-(ii)

Yield Example R1 R2 (%) Rf (Solvent) ¹H-NMR (400 MHz, δ) 2-1  H H 800.25 (CDCl₃): 1.10 (s, 9H), (MeOH:CH₂Cl₂ = 1:4) 1.84-1.89 (m, 4H), 3.07-3.13 (m, 2H), 3.38- 3.44 (m, 2H), 4.24 (s, 2H), 5.15 (s, 2H), 6.36 (s,1H), 6.73 (d, 1H), 7.10 (t, 1H), 7.22 (t, 1H), 7.47 (d, 1H), 8.84 (s,1H) 2-2  H

22 0.40 (MeOH:CH₂Cl₂ = 1:4) (CDCl₃): 1.10 (s, 9H), 1.90-1.95 (m, 2H),2.30- 2.50 (m, 2H), 2.65 (brs, 3H), 2.95-3.35 (m, 4H), 4.24 (s, 2H),5.14 (s, 2H), 6.36 (s, 1H), 6.74 (d, 1H), 7.13 (t, 1H), 7.22 (t, 1H),7.43 (d, 1H), 8.84 (s, 1H) 2-3  H

14 0.40 (MeOH:CH₂Cl₂ '2 1:4) (CDCl₃): ): 0.97 (t, 3H), 1.10 (s, 9H),1.90-1.95 (m, 2H), 2.30-2.50 (m, 2H), 2.95-3.35 (m, 6H), 4.25 (s, 2H),5.13 (s, 2H), 6.75 (d, 1H), 7.16 (t, 1H), 7.25 (t, 1H), 7.46 (d, 1H),8.85 (s, 1H) 2-4  H

27 0.45 (MeOH:CH₂Cl₂ = 1:4) (CDCl₃): 1.10 (s, 9H), 1.24 (d, 6H),1.92-2.00 (m, 2H), 2.15-2.30 (m, 2H), 2.90-3.13 (m, 3H), 3.15-3.25 (m,2H), 4.24 (s, 2H), 5.14 (s, 2H), 6.35 (s, 1H), 6.72 (d, 1H), 7.11 (t,1H), 7.20 (t, 1H), 7.46 (d, 1H), 8.83 (s, 1H) (s, 1H) 2-5  H

41 0.45 (MeOH:CH₂Cl₂ = 1:4) (CDCl₃): 0.97 (t, 3H), 1.10 (s, 9H),1.65-1.71 (m, 2H), 1.95-2.00 (m, 2H), 2.20-2.25 (m, 2H), 2.60-2.64 (m,2H), 2.90-3.00 (m, 2H), 3.05-3.15 (m, 2H), 4.24 (s, 2H), 5.14 (s, 2H),6.36 (s, 1H), 6.74 (d, 1H), 7.11 (t, 1H), 7.20 (t, 1H), 7.75 (d, 1H),8.84 (s, 1H) 2-6  H

33 0.15 (AcOEt only) (CDCl₃): 1.11 (s, 9H), 1.90-3.10 (m, 16H), 3.70-3.80 (m, 4H), 4.25 (s, 2H), 5.16 (s, 2H), 6.37 (s, 1H), 6.74 (d, 1H),7.31 (t, 1H), 7.24 (t, 1H), 7.45 (d, 1H), 8.85 (s, 1H) 2-7  H

60 0.55 (MeOH:CH₂Cl₂ = 1:4) (CDCl₃): 1.10 (s, 9H), 1.95-2.05 (m, 4H),2.75- 2.85 (m, 2H), 2.90- 3.05 (m, 2H), 3.67 (brs, 2H), 4.23 (s, 2H),5.14 (s, 2H), 6.35 (s, 1H), 6.72 (d, 1H), 7.02 (t, 1H), 7.09 (t, 1H),7.21 (t, 1H), 7.35 (t, 2H), 7.43 (d, 1H), 8.83 (s, 1H) 2-8  H

45 0.25 (AcOEt only) (CDCl₃): 1.10 (s, 9H) 1.55-1.59 (m, 2H), 1.86- 1.91(m, 4H), 2.18 (s, 3H), 3.73-3.79 (m, 2H), 4.24 (s, 2H), 5.16 (s, 2H),6.75 (d, 1H), 7.12 (t, 1H), 7.24 (t, 1H), 7.30 (d, 1H), 8.85 (s, 1H)2-9 

16 0.40 (MeOH:CH₂Cl₂ = 1:4) (CDCl₃): 1.09 (s, 9H), 1.93-2.10 (m, 4H),2.50 (s, 3H), 2.75-2.85 (m, 2H), 2.90-3.10 (m, 2H), 3.77 (s, 3H), 4.22(s, 2H), 5.12 (s, 2H), 6.36 (s, 1H), 6.62 (d,1H), 6.73 (dd, 1H), 7.02(d, 1H), 8.84 (s, 1H) 2-10

37 0.23 (MeOH:CH₂Cl₂ = 1:3) (DMSO): 1.04 (s, 9H), 1.07 (t, 3H),1.82-1.90 (m, 4H), 2.44-2.53 (m, 2H), 2.6- 2.69 (m, 2H), 2.78-2.87 (m,2H), 3.72 (s, 3H), 4.27 (s, 2H), 5.23 (s, 2H), 6.43 (s, 1H), 6.79 (d,1H), 6.83 (d, 1H), 7.15 (s, 1H), 9.02 (s, 1H), 2-11

H 43 0.10 (n-hexane:AcOEt = 1:1) (CDCl₃): 1.10 (s, 9H), 1.90-2.05 (m,2H), 2.30- 2.50 (m, 2H), 3.45- 3.60 (m, 2H), 3.90- 4.05 (m, 2H), 4.26(s, 2H), 5.15 (s, 2H), 6.36 (s, 1H), 6.72 (dd, 1H), 6.99 (td, 1H), 7.12(dd, 1H), 8.90 (s, 1H) 2-12

38 0.40 (MeOH:CH₂Cl₂ = 1:4) (CDCl₃): 1.09 (s, 9H), 1.92-2.10 (m, 4H),2.52 (s, 3H), 2.85-2.90 (m, 2H), 3.00-3.10 (m, 2H), 4.22 (s, 2H),5.13(s, 2H), 6.34 (s, 1H), 6.65 (dd, 1H), 6.92 (td, 1H), 7.16 (dd, 1H),8.85 (s, 1H) 2-13

27 0.38 (MeOH:CH₂Cl₂ = 1:5) (DMSO): 1.04 (s, 9H), 1.07 (t, 3H),1.82-1.95 (m, 4H), 2.44-2.53 (m, 2H), 2.6- 2.7 (m, 2H), 2.77-2.87 (m,2H), 4.27 (s, 2H), 5.26 (s, 2H), 6.44 (s, 1H), 6.92- 6.95 (m, 1H),7.04-7.09 (m, 1H), 7.47-7.49 (m, 1H), 9.01 (s, 1H),

Example 3 Example 3-0 Preparation of7-(2,2-Dimethyl-propyl)-6-(1,3-dioxo-2,8-diaza-spiro[4.5]dec-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

A. Preparation of 8-Benzyl-2,8-diaza-spiro[4.5]decane-1,3-dione

To a solution of 1-benzyl-piperidin-4-one (75.1 g, 0.40 mol) in toluene(400 ml), cyano-acetic acid ethyl ester (50.6 ml, 0.48 mol) and aceticacid (18.2 ml, 0.32 mol) are added at ambient temperature. The reactionmixture is refluxed for 4 h, quenched with ice-water and extracted withdiethyl ether. The combined extracts are washed with H₂O, brine anddried over sodium sulphate to give(1-benzyl-piperidin-4-ylidene)-cyano-acetic acid ethyl ester inquantitative yield.

Rf=0.53 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.30-1.37 (m, 3H), 2.58 (dd, 2H), 2.64 (dd,2H), 2.79 (dd, 2H), 3.15 (dd, 2H), 3.55 (s, 2H), 4.23-4.32 (m, 2H),7.21-7.36 (m, 5H).

To a solution of (1-benzyl-piperidin-4-ylidene)-cyano-acetic acid ethylester (112.9 g, 0.40 mol) in EtOH (500 ml) and H₂O (100 ml), potassiumcyanide (64.6 g, 0.99 mol) is added at ambient temperature. The reactionmixture is stirred at 65 C.° for 24 h. After removal of EtOH, H₂O isadded to the residue. The waster phase is extracted with diethyl ether.The combined extracts are washed with H₂O and brine, dried over sodiumsulfate and evaporated down to give 77.7 g of1-benzyl-4-cyanomethyl-piperidine-4-carbonitrile.

Rf=0.38 (n-hexane:AcOEt=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.76-1.81 (m, 2H), 2.10-2.05 (m, 2H),2.23-2.39 (m, 2H), 2.69 (s, 2H), 2.90-2.94 (m, 2H), 3.56 (s, 2H),7.21-7.38 (m, 5H).

Acetic acid (56.8 ml) and sulfuric acid (11.8 ml) are added to1-benzyl-4-cyanomethyl-piperidine-4-carbonitrile (27.2 g, 0.114 mmol) atambient temperature. The reaction mixture is stirred at 125 C.° for 1 h,cooled down to the room temperature and added to saturated NaOH aq. toadjust to pH 6.0. The mixture is extracted with dichloromethane. Thecombined extracts are washed with H₂O and brine, dried over sodiumsulfate and evaporated down to give8-benzyl-2,8-diaza-spiro[4.5]decane-1,3-dione (three steps yield:81.8%).

Rf=0.40 (CH₂Cl₂:MeOH=10:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.52-1.57 (m, 2H), 2.02-2.17 (m, 4H), 2.59(s, 2H), 2.86-2.90 (m, 2H), 3.52 (s, 2H), 7.21-7.28 (m, 2H), 7.30-7.37(m, 3H), 7.92 (brs, 1H).

B. Preparation of 1,3-Dioxo-2,8-diaza-spiro[4.5]decane-8-carboxylic acidtert-butyl ester

To 8-benzyl-2,8-diaza-spiro[4.5]decane-1,3-dione (28.3 g, 0.11 mol) andPd(OH)₂ (8.5 g) in 2 l of flask, EtOH (438 ml) and acetic acid (5.5 ml)are added at ambient temperature. The reaction mixture is stirred underH₂ at room temperature for 15 h. The catalysts are removed by filtrationand EtOH is evaporated down to give 2,8-diaza-spiro[4.5]decane-1,3-dionein quantitative yield. To a suspension of2,8-diaza-spiro[4.5]decane-1,3-dione (4.2 g, 25.2 mmol) indichloromethane (60 ml), 1N NaOH (26 ml, 26 mmol) anddi-t-butyldicarbonate (6.1 g, 27.7 mmol) in dichloromethane (20 ml) areadded at ambient temperature. The reaction mixture is stirred for 15 h.10% Citric acid is added to the reaction mixture and the pH of themixture is adjusted to 5. The combined extracts are washed with brine,dried over magnesium sulfate and concentrated under vacuum to give solidproduct, which filtrated with diethyl ether.

Yield: 51%

Rf=0.25 (n-hexane:ethyl acetate=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.47 (s, 9H), 1.55-1.70 (m, 2H), 1.95-2.05(m, 2H), 2.62 (s, 2H), 2.96-3.02 (m, 2H), 4.02-4.04 (m, 2H), 8.14 (brs,1H).

C. Preparation of2-[2-Cyano-7-(2,2-dimethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-1,3-dioxo-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester

6-Bromomethyl-7-(2,2-dimethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile(1.0 g, 3.25 mmol) and 1,3-dioxo-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester (0.82 g, 3.42 mmol) are dissolved in DMF (15 ml)and potassium carbonate (0.58 g, 4.23 mmol) is added to the solution.The reaction mixture is stirred at room temperature for 15 h andquenched with saturated ammonium chloride and extracted with ethylacetate. The combined extracts are washed with H₂O, brine and dried overmagnesium sulfate. Chromatography on silica gel (eluent; n-hexane:ethylacetate=2:1) gives 1.56 g of desired2-[2-cyano-7-(2,2-dimethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-1,3-dioxo-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester in 97% yield.

Rf=0.30 (n-hexane:ethyl acetate=1:1).

¹H-NMR (400 MHz, CDCl₃) δ: 0.99 (s, 9H), 1.40 (s, 9H), 1.66-1.68 (m,4H), 2.89-2.93 (m, 2H), 3.85-3.88 (m, 2H), 4.25 (s, 2H), 4.90 (s, 2H),6.62 (s, 1H), 9.06 (s, 1H).

D. Preparation of7-(2,2-Dimethyl-propyl)-6-(1,3-dioxo-2,8-diaza-spiro[4.5]dec-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

To a solution of2-[2-cyano-7-(2,2-dimethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-1,3-dioxo-2,8-diaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester (1.5 g, 3.1 mmol) in dichloromethane (20 ml), TFA(5 ml) is added at 0° C. The reaction mixture is stirred at roomtemperature for 2 h. After removal of the solvent, saturated sodiumbicarbonate is added to the residue and the mixture is extracted withdichloromethane. The combined extracts are washed with H₂O, brine, driedover magnesium sulfate and concentrated under vacuum to give desiredproduct,7-(2,2-dimethyl-propyl)-6-(1,3-dioxo-2,8-diaza-spiro[4.5]dec-2-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile.

Yield: 91%.

Rf=0.15 (CH₂Cl₂:MeOH=10:1).

¹H-NMR (400 MHz, CDCl₃) δ: 1.03 (s, 9H), 1.4-1.51 (m, 2H), 1.69 (brs,1H), 1.95-2.02 (m, 2H), 2.66 (s, 2H), 2.69-2.72 (m, 2H), 3.11-3.17 (m,2H), 4.34 (s, 2H), 4.91 (s, 2H), 6.59 (s, 1H), 8.90 (s, 1H).

By repeating the procedures described above using appropriate startingmaterials and conditions the following compounds of formula I-(iii) areobtained as identified below in Table 3.

TABLE 3 Formula I-(iii)

Example Rx Yield (%) Rf (Solvent) NMR (400 MHz, δ) 3-1

41 0.55 (MeOH:CH₂Cl₂ = 1:10) (CDCl₃): 1.02 (s, 9H), 1.06- 1.16 (m, 3H),1.31-1.42 (m, 2H), 1.52-1.72 (m, 6H), 1.92-2.02 (m, 3H), 2.64 (s, 2H),2.98-3.05 (m, 2H), 3.61-3.65 (m, 1H), 3.84-3.90 (m, 2H), 4.28-4.30 (m,1H), 4.33 (s, 2H), 4.92 (s, 2H), 6.60 (s, 1H), 8.90 (s, 1H) 3-2

46 0.50 (MeOH:CH₂Cl₂ = 1:10) (CDCl₃): 1.02 (s, 9H), 1.53- 1.56 (m, 2H),1.97-2.03 (m, 2H), 2.65 (s, 2H), 2.81 (d, 3H), 3.01-3.08 (m, 2H),3.86-3.91 (m, 2H), 4.33 (s, 2H), 4.52- 4.52 (m, 1H), 4.92 (s, 2H), 6.60(s, 1H), 8.90 (s, 1H) 3-3

28 0.30 (n-hexane:AcOEt = 1:1) (CDCl₃): ): 1.03 (s, 9H), 1.55-1.68 (m,3H), 2.29- 2.36 (m, 2H), 2.60-2.68 (m, 3H), 3.36-3.40 (m, 2H), 3.77 (s,3H), 3.84 (s, 3H), 4.34 (s, 2H), 4.93 (s, 2H), 6.41 (d, 1H), 6.45 (s,1H), 6.49 (s, 1H), 6.85 (d, 1H), 8.90 (s, 1H) 3-4

54 0.45 (MeOH:CH₂Cl₂ = 1:10) (CDCl₃): 1.02 (s, 9H), 1.52- 1.68 (m, 2H),1.94- 2.06 (m, 3H), 2.11 (s, 3H), 2.68 (d, 2H), 3.05-3.15 (m, 1H),3.25-3.35 (m, 1H), 3.83-3.95 (m, 1H), 4.28-4.36 (m, 3H), 4.93 (s, 2H),6.61 (s, 1H), 8.90 (s, 1H) 3-5

62 0.35 (n-hexane:AcOEt = 1:1) (CDCl₃): 1.02 (s, 9H), 1.45- 1.50 (m,2H), 2.00- 2.14 (m, 4H), 2.60 (s, 2H), 2.86-2.90 (m, 2H), 3.52 (s, 2H),4.32 (s, 2H), 4.90 (s, 2H), 6.58 (s, 1H), 7.25-7.32 (m, 5H), 8.89 (s,1H) 3-6

30 0.20 (n-hexane:AcOEt = 1:1) (CDCl3): 1.03 (s, 9H), 1.55-1.60 (m, 2H),1.90- 2.10 (m, 2H), 2.45-2.55 (m, 4H), 2.68 (brs, 2H), 3.09-3.31 (m,4H), 3.70- 3.72 (m, 4H), 4.09-4.14 (m, 2H), 4.34 (s, 2H), 4.93 (s, 2H),6.61 (s, 1H), 8.90 (s, 1H)

Example 4 Example 4-0 Preparation of6-(8-Acetyl-2,8-diaza-spiro[4.5]dec-2-ylmethyl)-7-(3,3-dimethyl-butyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

8-Methanesulfonyl-2,8-diaza-spiro[4.5]decane hydrochloride

To a solution of 2,8-diaza-spiro[4.5]decane-2-carboxylic acid tert-butylester (1.12 g, 4.66 mol) in CH₂Cl₂ (10 ml), triethylamine (3.88 ml) andmethanesulfonyl chloride (1.08 ml, 14 mmol) are added at 0° C. Thereaction mixture is stirred for overnight, quenched with ice-water andextracted with dichloromethane. The combined extracts are washed withH₂O, brine and dried over sodium sulphate to give crude 8-methanesulfonyl-2,8-diaza-spiro[4.5]decane-2-carboxylic acid tert-butyl ester(1.32 g).

Rf=0.7 (CH₂Cl₂:MeOH=10:1). ¹H-NMR (400 MHz, CDCl₃) δ: 1.46 (s, 9H),1.66-1.76 (m, 6H), 2.76-2.80 (m, 2H), 3.00 (s, 3H), 3.15-3.25 (m, 2H),3.36-3.45 (m, 4H).

To a solution of8-methanesulfonyl-2,8-diaza-spiro[4.5]decane-2-carboxylic acidtert-butyl ester (1.32 g) in ethyl acetate (10 ml), a 1 M ethyl acetatesolution of HCl (20 ml). After stirring for 2 h at room temperature,solvent is evaporated down to give8-methanesulfonyl-2,8-diaza-spiro[4.5]decane hydrochloride as a solid.

Rf=0.05 (ethyl acetate only). ¹H-NMR (400 MHz, DMSO-d₆) δ: 1.62-1.68 (m,4H), 1.78-1.82 (m, 2H), 2.87 (s, 3H), 2.98-3.12 (m, 6H), 3.20-3.23 (m,2H), 9.49 (brs, 1H), 9.59 (brs, 1H).

1-(2,8-Diaza-spiro[4.5]dec-8-yl)-ethanone hydrochloride

To a solution of 2,8-diaza-spiro[4.5]decane-2-carboxylic acid tert-butylester (1.12 g, 4.66 mol) in dichloromethane (10 ml), triethylamine (3.88ml) and acetic anhydride (1.32 ml, 14 mmol) are added at 0° C. Thereaction mixture is stirred for overnight, quenched with ice-water andextracted with dichloromethane. The combined extracts are washed withH₂O, brine and dried over sodium sulphate to give crude8-acetyl-2,8-diaza-spiro[4.5]decane-2-carboxylic acid tert-butyl ester(1.34 g).

Rf=0.6 (CH₂Cl₂:MeOH=10:1) ¹H-NMR (400 MHz, CDCl₃) δ: 1.46 (s, 9H),1.50-1.56 (m, 4H), 1.72-1.76 (m, 2H), 2.03 (s, 2H), 2.22 (s, 3H),3.16-3.49 (m, 6H).

To a solution of 8-acetyl-2,8-diaza-spiro[4.5]decane-2-carboxylic acidtert-butyl ester (1.34 g) in ethyl acetate (10 ml), a 1 M ethyl acetatesolution of HCl (20 ml). After stirring for 2 h at room temperature, thereaction mixture is evaporated down to give1-(2,8-diaza-spiro[4.5]dec-8-yl)-ethanone hydrochloride as a solid.

Rf=0.05 (ethyl acetate only). ¹H-NMR (400 MHz, DMSO-d₆) δ: 1.44-1.59 (m,4H), 1.76-1.83 (m, 2H), 2.07 (s, 3H), 2.96-3.06 (m, 2H), 3.16-3.24 (m,4H), 3.38-3.56 (m, 2H), 9.55 (brs, 1H), 9.67 (brs, 1H).

Intermediate I

To a solution of 5-methoxy-1,3-dihydro-indol-2-one (1.5 g, 10 mmol) inTHF (160 ml), a solution of NaHMDS (1 M THF solution) (50 ml, 50 mmol)is added at −78° C. After stirring for 30 min at −78° C.,ethyl-bis-(2-chloro-ethyl)-amine (47.3 g, 0.18 mol) in THF (176 ml) isadded and the reaction mixture is stirred for 15 h at room temperature,quenched with sat.NH₄Claq. and ice-water and extracted with ethylacetate. The combined extracts are washed with brine, dried over sodiumsulphate and evaporated down. Ethyl ether is added to the residue togive the powder, which is filtrated.

Rf=0.10 (CH₂Cl₂:MeOH=30:1) ¹H-NMR 400 MHz, CDCl₃) δ: 1.17 (t, 3H),1.87-2.02 (m, 4H), 2.60 (q, 2H), 2.69-2.74 (m, 2H), 2.90-2.96 (m, 2H),6.78-6.82 (m, 1H), 6.88-6.93 (m, 1H), 7.08-7.11 (m, 1H), 8.04 (brs, 1H).

Intermediate L

To a solution of 5-Methoxy-1,3-dihydro-indol-2-one (1.06 g, 6.49 mmol)in THF (13 ml), a solution of NaHMDS (1 M THF solution) (32.5 ml, 32.5mmol) is added at −78° C. After stirring for 30 min at −78° C.,methyl-bis-(2-chloro-ethyl)-amine hydrochloride (1.37 g, 7.14 mol) isadded and the reaction mixture is stirred for 13.5 h at roomtemperature, quenched with sat.NH₄Claq. and ice-water and extracted withethyl acetate. The organic extracts are washed with brine, dried oversodium sulphate and evaporated down. Ethyl ether is added to the residueto give the powder, which is filtrated.

Rf=0.10 (CH₂Cl₂:MeOH=30:1) ¹H-NMR (400 HMz, DMSO-d₆) δ: 1.66-1.78 (m,4H), 2.28 (s, 3H), 2.44-2.47 (m, 2H), 2.71-2.77 (m, 2H), 3.70 (s, 3H),6.74 (s, 2H), 7.01 (s, 1H), 10.15 (brs, 1H).

Intermediate

To a solution of intermediate (422 mg, 1.76 mol) in dichloromethane (5ml), triethylamine (1.2 ml) and acetic anhydride (0.33 ml, 3.53 mmol)are added at 0° C. The reaction mixture is stirred for 2 h, and isquenched with ice-water and extracted with dichloromethane. The combinedorganic layer is washed with water and brine, dried over MgSO₄, andconcentrated in vacuo. The residue is purified by silica gel columnchromatography (n-hexane:AcOEt=5:1) to give the product.

Rf=0.6 (CH₂Cl₂:MeOH 10:1) ¹H-NMR (400 MHz, CDCl₃) δ: 1.79-1.95 (m, 4H),2.20 (s, 3H), 3.68-3.74 (m, 1H), 3.80-3.87 (m, 1H), 3.98-4.22 (m, 2H),6.90-6.92 (m, 1H), 7.03-7.07 (m, 1H), 7.22-7.26 (m, 2H), 8.06 (brs, 1H).

6-(8-Acetyl-2,8-diaza-spiro[4.5]dec-2-ylmethyl)-7-(3,3-dimethyl-butyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

To a solution of6-bromomethyl-7-(3,3-dimethyl-butyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile(40 mg, 1.37 mmol) in DMF (5 ml),1-(2,8-diaza-spiro[4.5]dec-8-yl)-ethanone hydrochloride (300 mg, 1.37mmol) and K₂CO₃ (568 mg, 4.11 mmol) and triethylamine (5 ml) are added.The mixture is stirred at room temperature under nitrogen atmosphere for11 h. The reaction mixture is diluted with water and extracted withAcOEt (twice). The combined organic layers are washed with water andbrine, dried over MgSO₄, and concentrated in vacuo. The residue ispurified by silica gel column chromatography (n-hexane:AcOEt=1:1) togive the product.

Rf=0.30 (n-hexane:AcOEt=1:1). ¹H-NMR (400 MHz, CDCl₃) δ: 1.05 (s, 9H),1.53-1.72 (m, 8H), 2.07 (s, 3H), 2.40-2.48 (m, 2H), 2.60-2.69 (m, 2H),3.35-3.45 (m, 2H), 3.60-3.67 (m, 1H), 3.74-3.82 (m, 2H), 4.40-4.44 (m,2H), 6.49 (s, 1H), 8.87 (s, 1H).

By repeating the procedures described above using appropriate startingmaterials and conditions the following compounds of formula I-iv areobtained as identified below in Table 4.

TABLE 4 Formula 1-iv

Example R3 R4 Rf (solvent) NMR (400 MHz, CDCl3, □) 4-1 

0.80 (n-hexane: AcOEt = 1:1) 1.50-1.71 (m, 6H), 2.06 (s, 3H), 2.32-2.41(m, 2H), 2.48-2.65 (m, 2H), 3.10- 3.14 (m, 2H), 3.29-3.52 (m, 5H),3.62-3.69 (m, 1H), 4.58-4.61 (m, 2H), 6.46 (s, 1H), 6.99-7.01 (m, 2H),7.23-7.26 (m, 2H), 8.89 (s, 1H). 4-2 

0.70 (n-hexane: AcOEt = 1:1) 1.53-1.55 (m, 2H), 1.63- 1.70 (m, 6H), 2.35(s, 2H), 2.56-2.60 (m, 2H), 2.75 (s, 3H), 3.05-3.13 (m, 2H), 3.20-3.26(m, 2H), 3.46 (s, 2H), 4.57-4.61 (m, 2H), 6.45 (s, 1H), 6.97-6.99 (m,2H), 7.22-7.25 (m, 2H), 8.90 (s, 1H). 4-3 

0.80 (n-hexane: AcOEt = 1:1) 1.04 (s, 9H), 1.66-1.70 (m, 8H), 2.43 (brs,2H), 2.62- 2.65 (m, 2H), 2.75 (s, 3H), 3.09-3.15 (m, 2H), 3.20- 3.25 (m,2H), 3.78 (s, 2H), 4.39-4.43 (m, 2H), 6.49 (s, 1H), 8.88 (s, 1H). 4-4 

0.30 (n-hexane: AcOEt = 1:1) 0.97-1.03 (m, 2H), 1.15- 1.34 (m, 5H),1.56-1.80 (m, 12H), 2.35-2.40 (m, 6H), 2.55-2.58 (m, 2H), 3.45 (s, 2H),3.75 (s, 2H), 4.38- 4.41 (m, 2H), 6.47 (s, 1H), 7.29-7.30 (m, 5H), 8.86(s, 1H). 4-5 

0.13 (n-hexane: AcOEt = 1:1) 1.53-1.60 (m, 4H), 2.09- 2.16 (m, 4H), 2.59(s, 2H), 2.80-2.83 (m, 2H), 3.12- 3.14 (m, 2H), 3.37 (s, 2H), 4.55-4.64(m, 2H), 6.47 (s, 1H), 6.99-7.03 (m, 2H), 7.23-7.26 (m, 2H), 7.75 (brs,1H), 8.90 (s, 1H). 4-6 

0.10 (n-hexane: AcOEt = 1:1) 0.98-1.39 (m, 9H), 1.65- 1.82 (m, 7H),1.99-2.03 (m, 4H), 2.59-2.64 (m, 2H), 2.74-2.77 (m, 2H), 2.92- 2.98 (m,2H), 4.36-4.39 (m, 2H), 5.10 (s, 2H), 6.40 (s, 1H), 6.69-6.72 (m, 1H),6.88-6.93 (m, 1H), 7.16- 7.18 (m, 1H), 8.86 (s, 1H). 4-7 

0.10 (n-hexane: AcOEt = 1:1) 0.97-1.39 (m, 6H), 1.60- 1.82 (m, 8H),1.98-2.00 (m, 3H), 2.46 (s, 3H), 2.71- 2.74 (m, 2H), 2.92-2.94 (m, 2H),3.77 (s, 3H), 4.36- 4.40 (m, 2H), 5.09 (s, 2H), 6.40 (s, 1H), 6.66-6.73(m, 2H), 7.02 (d, 1H), 8.85 (s, 1H). 4-8 

0.30 (n-hexane: AcOEt = 1:1) 1.02-1.42 (m, 6H), 1.68- 1.95 (m, 1H), 2.12(s, 3H), 3.75-3.85 (m, 2H), 4.01- 4.07 (m, 1H), 4.24-4.29 (m, 1H),4.40-4.44 (m, 2H), 5.16 (s, 2H), 6.44 (s, 1H), 6.84-6.86 (m, 1H), 7.11-7.15 (m, 1H), 7.24-7.33 (m, 2H), 8.85 (s, 1H). 4-9 

0.35 (n-hexane: AcOEt = 1:1) 1.09 (s, 9H), 1.70-1.74 (m, 2H), 1.88-1.94(m, 4H), 2.19 (s, 3H), 3.74-3.81 (m, 2H), 4.04-4.14 (m, 1H), 4.26-4.29(m, 1H), 4.38- 4.42 (m, 2H), 5.13 (s, 2H), 6.38 (s, 1H), 6.80 (d, 1H),7.11-7.15 (m, 1H), 7.23- 7.32 (m, 2H), 8.85 (s, 1H). 4-10

0.30 (n-hexane: AcOEt = 1:1) 1.38-1.93 (m, 13H), 2.08- 2.17 (m, 2H),2.19 (s, 3H), 3.72-3.84 (m, 2H), 3.99- 4.06 (m, 1H), 4.23-4.29 (m, 1H),4.41-4.45 (m, 2H), 5.12 (s, 2H), 6.48 (s, 1H), 6.84-6.86 (m, 1H),7.11-7.15 (m, 1H), 7.24-7.32 (m, 2H), 8.89 (s, 1H) 4-11

0.20 (n-hexane: AcOEt = 1:1) (DMSO-d₆) 1.07 (t, 3H), 1.24-1.46 (m, 3H),1.69-2.02 (m, 12H), 2.60-2.75 (m, 2H), 2.80- 2.90 (m, 2H), 3.25-3.36 (m,2H), 4.40-4.44 (m, 2H), 5.26 (s, 2H), 6.54 (s, 1H), 7.04-7.09 (m, 2H),7.22- 7.25 (m, 1H), 7.55-7.75 (m, 1H), 9.02 (s, 1H). 4-12

0.09 (n-hexane: AcOEt = 1:1) CDCl3 0.98-1.07 (m, 2H), 1.18- 1.41 (m,4H), 1.68-1.84 (m, 9H), 2.11-2.16 (m, 2H), 2.27-2.32 (m, 2H), 2.92- 2.99(m, 2H), 3.03 (s, 3H), 3.73-3.78 (m, 2H), 4.40- 4.44 (m, 2H), 5.84 (brs,1H), 6.53 (s, 1H), 8.89 (s, 1H)

Example 4-13 Preparation of3-[2-Cyano-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrmidin-6-ylmethyl]-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester

4-Oxo-1-phenyl-1,3,8-triaza-spiro[4.5]decane-8-carboxylic acidtert-butyl ester

To a suspension of 1-phenyl-1,3,8-triaza-spiro[4.5]decan-4-one (1.0 g,4.32 nmol) in dichloromethane (10 ml), saturated sodium bicarbonatesolution (10 ml) and di-t-butyldicarbonate (1.04 g, 4.76 mmol) indichloromethane (5 ml) are added at ambient temperature. The reactionmixture is stirred for 1 h and quenched with H₂O and extracted withethyl acetate. The combined extracts are washed with H₂O and brine,dried over sodium sulfate and evaporated down to give4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]decane-8-carboxylic acidtert-butyl ester. Yield: 100%

Rf=0.90 (CH₂Cl₂:MeOH=20:1) ¹H-NMR (400 MHz, CDCl₃) δ: 1.51 (s, 9H),1.63-1.71 (m, 2H), 2.50-2.65 (m, 2H), 3.50-3.65 (m, 2H), 3.97-4.10 (m,2H), 4.75 (s, 2H), 6.74-6.76 (m, 2H), 6.84-6.88 (m, 1H), 7.01 (brs, 1H),7.23-7.27 (m, 2H).

3-[2-Cyano-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester

To a solution of6-chloromethyl-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile(600 mg, 1.98 mmol) in DMF (7 ml),4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]decane-8-carboxylic acidtert-butyl ester (657 mg, 1.98=mol) and sodium hydride (101 mg, 2.53mmol) are added. The mixture is stirred at room temperature undernitrogen atmosphere for 14 h. The reaction mixture is diluted with waterand extracted with AcOEt (twice). The combined organic layers are washedwith water and brine, dried over MgSO₄, and concentrated in vacuo. Theresidue is purified by silica gel column chromatography(n-hexane:AcOEt=1:1) to give the product in 29% yield.

Rf=0.25 (n-hexane:AcOEt=1:1). ¹H-NMR (400 MHz CDCl₃) δ: 0.97-1.49 (m,7H), 1.50 (s, 9H), 1.56-1.82 (m, 8H), 2.45-2.60 (m, 2H), 3.50-3.65 (m,2H), 4.09-4.14 (m, 2H), 4.33-4.36 (m, 2H), 4.64 (s, 2H), 4.87 (s, 2H),6.72-6.74 (m, 2H), 6.86-6.90 (m, 1H), 7.20-7.24 (m, 2H), 8.94 (s, 1H).

Example 4-147-(2-Cyclohexyl-ethyl)-6-(4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitriletrifluoroacetic acid salt

To a solution of3-[2-cyano-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]decane-8-carboxylicacid tert-butyl ester (340 mg, 0.56 mmol) in dichloromethane (5 ml),trifluoroacetic acid (5 ml) is added. After stirring for 1 h at roomtemperature, solvent is evaporated down to give7-(2-cyclohexyl-ethyl)-6-(4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitriletrifluoroacetic acid salt in quant yield.

Rf=0.10 (CH₂Cl₂:MeOH=20:1) ¹H-NMR (400 MHz, CDCl₃) δ: 0.98-1.38 (m, 5H),1.65-1.83 (m, 8H), 1.98-2.09 (m, 2H), 2.71-2.80 (m, 2H), 3.53-3.56 (m,2H), 3.94-4.02 (m, 2H), 4.38-4.42 (m, 2H), 4.73 (s, 2H), 4.91 (s, 2H),6.71 (s, 1H), 6.88-6.90 (m, 2H), 7.01-7.04 (m, 1H), 7.28-7.32 (m, 2H),7.85 (brs, 1H), 8.25 (brs, 1H), 9.08 (s, 1H).

Example 4-156-(8-Acetyl-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-ylmethyl)-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

To a solution of7-(2-cyclohexyl-ethyl)-6-(4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitriletrifluoroacetic acid salt (142 mg, 0.28 mol) in dichloromethane (2 ml),triethylamine (395 μl) and acetic anhydride (54 μl, 0.57 mmol) are addedat 0° C. The reaction mixture is stirred for over night at roomtemperature, quenched with ice-water and extracted with ethyl acetate.The combined extracts are washed with H₂O, brine and dried over sodiumsulphate. Chromatography on silica gel gives 90 mg of6-(8-acetyl-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-ylmethyl)-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrilein 58% yield.

Rf=0.30 (n-hexane:AcOEt=1:1). ¹H-NMR (400 MHz CDCl₃) δ: 0.97-1.40 (m,6H), 1.64-1.82 (m, 9H), 2.14 (s, 3H), 2.37-2.44 (m, 2H), 3.40-3.48 (m,1H), 3.74-3.79 (m, 1H), 3.93-4.01 (m, 1H), 4.34-4.38 (m, 2H), 4.56-4.66(m, 3H), 4.87 (s, 2H), 6.61 (s, 1H), 6.74-6.76 (m, 2H), 6.91-6.95 (m,1H), 7.23-7.25 (m, 2H), 8.94 (s, 1H).

Example 4-166-(2-Acetyl-2,8-diaza-spiro[4.5]dec-8-ylmethyl)-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile

To a solution of6-bromomethyl-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitrile(290 mg, 0.84 mmol) in DMF (1.7 ml),2,8-Diaza-spiro[4.5]decane-2-carboxylic acid tert-butyl ester (201 mg,0.84 mmol) and potassium carbonate (138 mg, 1.0 mmol) are added. Themixture is stirred at room temperature under nitrogen atmosphere for 14h. The reaction mixture is diluted with water and extracted with AcOEt(twice). The combined organic layer is washed with water and brine,dried over MgSO₄, and concentrated in vacuo. The residue is purified bysilica gel column chromatography (n-hexane:AcOEt=1:1) to give8-[2-Cyano-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-2,8-diaza-spiro[4.5]decane-2-carboxylicacid tert-butyl ester in 71% yield. Rf=0.45 (n-hexane:AcOEt=1:1).

To a solution of8-[2-cyano-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-6-ylmethyl]-2,8-diaza-spiro[4.5]decane-2-carboxylicacid tert-butyl ester (300 mg, 0.59 mmol) in dichloromethane (5 ml),trifluoroacetic acid (3 ml) is added. After stirring for 1.5 h at roomtemperature, solvent is evaporated down to give7-(2-cyclohexyl-ethyl)-6-(2,8-diaza-spiro[4.5]dec-8-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitriletrifluoroacetic acid salt in quant yield.

Rf=0.10 (CH₂Cl₂:MeOH=10:1)

To a solution of7-(2-cyclohexyl-ethyl)-6-(2,8-diaza-spiro[4.5]dec-8-ylmethyl)-7H-pyrrolo[2,3-d]pyrimidine-2-carbonitriletrifluoroacetic acid salt in pyridine (5 ml), acetic anhydride (0.28 ml,2.90 mmol) are added at 0° C. The reaction mixture is stirred for overnight at room temperature, quenched with ice-water and extracted withethyl acetate. The combined extracts are washed with H₂O, brine anddried over sodium sulphate. Chromatography on silica gel gives 79 mg of6-(2-acetyl-2,8-diaza-spiro[4.5]dec-8-ylmethyl)-7-(2-cyclohexyl-ethyl)-7H-pyrrolo[2,3]pyrimidine-2-carbonitrilein 30% yield (3 steps).

Rf=0.30 (n-hexane:AcOEt=1:1). ¹H-NMR (400 MHz, CDCl₃) δ: 1.00-1.84 (m,17H), 2.04 (s, 3H), 2.33-2.56 (m, 4H), 3.25-3.35 (m, 2H), 3.47-3.53 (m,2H), 3.66-3.69 (m, 2H), 4.38-4.43 (m, 2H), 6.49 (s, 1H), 8.87 (s, 1H).

1. A compound of formula I, or a pharmaceutically acceptable salt orester thereof:

wherein E is a radical of formula a or formula b

wherein A is CH₂, CH₂—CH₂ or C═O;

B is CH₂, C═O or D is CH₂, or C═O; G is CH₂, CH₂C═O or CH₂—CH₂; J isCH₂, C═O or CH₂—CH₂; M is CH₂ or NH; Q is H, lower alkyl, hydroxysubstituted lower alkyl, optionally substituted aryl lower alkyl, loweralkyl sulfonyl, carbocyclic aryl lower alkyl, lower alkoxy-substitutedcarbocyclic aryl lower alkyl, halo-substituted carbocyclic aryl loweralkyl, N-heterocyclyl-substituted lower alkyl, lower alkoxy substitutedcarbocyclic aryl, amino carbonyl, cycloalkyl amino carbonyl,N-heterocyclyl substituted lower alkyl carbonyl, halo-substitutedcarbocyclic aryl lower alkyl, lower alkoxy carbonyl, or lower alkylcarbonyl; and R is lower alkyl, para-chlorophenylethyl, cyclohexylethyl,dimethylbutyl, difluorocyclohexylethyl, cyclopentylethyl orcycloheptylethyl.
 2. A compound of formula I-(i) or a pharmaceuticallyacceptable salt or ester thereof

wherein Q-i is H, lower alkyl, hydroxyl-substituted lower alkyl,N-heterocyclyl substituted lower alkyl, mono or di-substituted aryllower alkyl, lower alkoxy substituted carbocyclic aryl lower alkyl; andR is as defined in claim
 1. 3. (canceled)
 4. A compound of formulaI-(iii) or a pharmaceutically acceptable salt or ester thereof

wherein B-iii is CH₂; G-iii is CH₂CH₂; J-iii is CH₂CH₂; Q-iii is H,cycloalkyl amino carbonyl, amino carbonyl, lower alkoxy substitutedcarbocyclic aryl, lower alkyl carbonyl, carbocyclic aryl lower alkyl orN-heterocyclyl substituted lower alkyl carbonyl; and R is as defined inclaim
 1. 5. A compound of claim 1, wherein R is R1 which is lower alkyl.6. A compound of claim 1, wherein R is R2 which ispara-chlorophenylethyl, cyclohexylethyl, dimethylbutyl,difluorocyclohexylethyl, cyclopentylethyl or cycloheptylethyl.
 7. Acompound of claim 1, wherein R is R5 which is 2,2-dimethyl-propyl.
 8. Acompound of claim 1, wherein R is R6 which is 3,3-dimethyl-butyl.
 9. Acompound of formula I-(iv) or a pharmaceutically acceptable salt orester thereof

wherein R3 is (8-loweralkyl-carbonyl)-2,8-diaza-spiro[4.5]dec-2-ylmethyl, (8-loweralkyl-sulfonyl)-2,8-diaza-spiro[4.5]dec-2-ylmethyl, (8-aryl-loweralkyl)-2,8-diaza-spiro[4.5]dec-2-ylmethyl,

R4 is para-chlorophenylmethyl, cyclohexylmethyl, dimethylpropyl,difluorocyclohexylmethyl, cyclopentylmethyl or cycloheptylmethyl; and Qis as defined in claim
 1. 10-11. (canceled)
 12. A pharmaceuticalcomposition comprising an excipient and a compound according to claim 1as an active ingredient.
 13. A method of treating a patient sufferingfrom or susceptible to a disease or medical condition in which cathepsinK and/or cathepsin S is implicated, comprising administering aneffective amount of a compound according to claim 1 to the patient. 14.(canceled)
 15. A process for the preparation of a compound of claim 1comprising coupling a compound of formula II

wherein Q is defined in claim 1, with a compound of formula III

wherein X is a halo and R is defined in claim 1; and recovering theresulting compound in free base, or in a pharmaceutically acceptablesalt or ester thereof.